羊草叶绿体非编码区多态性标记筛选及群体遗传多样性

为筛选多态性丰富的羊草叶绿体非编码区片段,本研究以9个不同地理位置的羊草居群为材料,通过对12个叶绿体非编码区DNA片段测序及其序列间变异分析试图从中找出有遗传差异的叶绿体DNA(cpDNA)片段,并利用有多态性的cpDNA片段进行遗传多样性分析。结果表明,序列ndhF-rpl32、trnL-trnF、trnC-ycf6、aptI-aptH具有比较丰富的变异,可作为下一步研究野生羊草群体遗传学和谱系地理学较为理想的分子标记。在37个羊草个体4条非编码区片段的合并序列中,共检测到15种单倍型,单倍型多态性(Hd)为0.928,核苷酸多态性(Pi)为0.00101;遗传分化指数(Fst)为0.58884,基因流(Nm)为0.17,基因流较小;中性检验Tajima’s D(-1.08542)和Fu’s Fs(-5.301)均为负值,且差异不显著(P>0.10),推测羊草遵循中性进化理论,可能经历过种群扩张;AMOVA结果显示,65%的分子变异出现在居群间,35%出现在居群内;Mantel检验得到,遗传距离与地理距离具有显著相关性(r=0.449,P<0.05);居群分化值Nst 0.386(0.1865)大于Gst 0.234(0.1506),差异显著(P<0.05),表明羊草居群存在分子谱系地理结构。通过系统发育树分析得到,羊草15个单倍型分为两大分支,H2和H10聚为一支,它们与别的居群个体亲缘关系较远,其余单倍型聚为另一支;单倍型网络图显示,H2和H12、H10和H12亲缘关系较远,与系统发育树分析结果基本一致。本研究为羊草的种质资源保护、谱系地理学研究等工作奠定了基础。     

Ergovaline does not alter the severity of ryegrass staggers induced by lolitrem B

AIM: To investigate a possible interaction between lolitrem B and ergovaline by comparing the incidence and severity of ryegrass staggers in sheep grazing ryegrass (Lolium perenne) containing lolitrem B or ryegrass containing both lolitrem B and ergovaline.

METHODS: Ninety lambs, aged approximately 6 months, were grazed on plots of perennial ryegrass infected with either AR98 endophyte (containing lolitrem B), standard endophyte (containing lolitrem B and ergovaline) or no endophyte, for up to 42 days from 2 February 2010. Ten lambs were grazed on three replicate plots per cultivar. Herbage samples were collected for alkaloid analysis and lambs were scored for ryegrass staggers (scores from 0–5) weekly during the study. Any animal which was scored ≥4 was removed from the study.

RESULTS: Concentrations of lolitrem B did not differ between AR98 and standard endophyte-infected pastures during the study period (p=0.26), and ergovaline was present only in standard endophyte pastures. Ryegrass staggers was observed in sheep grazing both the AR98 and standard endophyte plots, with median scores increasing in the third week of the study. Prior to the end of the 42-day grazing period, 22 and 17 animals were removed from the standard endophyte and AR98 plots, respectively, because their staggers scores were ≥4. The cumulative probability of lambs having scores ≥4 did not differ between animals grazing the two pasture types (p=0.41).

CONCLUSIONS AND CLINICAL RELEVANCE: There was no evidence for ergovaline increasing the severity of ryegrass staggers induced by lolitrem B. In situations where the severity of ryegrass staggers appears to be greater than that predicted on the basis of concentrations of lolitrem B, the presence of other tremorgenic alkaloids should be investigated.     

青海湖高寒沙区引进辽西樟子松治沙研究

通过引进东北辽西科尔沁沙地樟子松在青海湖斜麻沙区进行栽植试验,结果表明,引进的樟子松当年主顶芽平均生长高度达10.13 cm,侧顶芽生长高度平均达5.14 cm,平均顶芽生长达7.64 cm,苗木整体生长良好,当年成活率达到93%;经过长达6个多月寒冬,越冬成活率达80%,返青良好。     

铺地竹叶片对结球生菜种子发芽及幼苗生长的影响

采用基质盆栽试验,研究了铺地竹(Sasa argenteastriatus E.G.Camus)叶片对结球生菜(Lactucasativa L.var.capitata L.)种子萌发和幼苗生长的影响,结果表明,铺地竹叶片粉末对结球生菜种子的萌发和幼根生长具有一定的抑制作用,随着浓度的增加抑制作用增强,在50～200 g/m2的范围内,幼根抑制率为12.15%～28.91%。在50～100 g/m2的用量下,铺地竹叶片对结球生菜苗长表现为促进生长作用,而在200 g/m2时,则表现为抑制生长作用。施用铺地竹叶片后3 d、6 d、9 d和12 d,结球生菜幼苗的叶绿素含量特别是叶绿素a的含量显著减少,随着基质中叶片粉末浓度的增加,结球生菜叶绿素含量降低,叶绿素a含量高低依次为对照、低浓度处理、中浓度处理、高浓度处理。随着处理后时间的延长,竹叶对结球生菜叶片叶绿素a含量的影响减弱。这个结果进一步验证了自然条件下铺地竹叶片对植物生长的化感效应,说明其具有开发植物源除草剂的潜力,为研究铺地竹叶片中的化感物质提供了佐证。     

青花菜锌指蛋白基因BoCCCH1的克隆和表达分析

CCCH锌指蛋白广泛存在于真核生物中,在植物的生长、发育和逆境胁迫响应中起着重要作用。为获知青花菜CCCH转录因子基因的序列特征和表达特性,从青花菜叶片中克隆到1个CCCH基因,命名为Bo CCCH1,同时,利用RT-PCR方法研究了其在不同器官及霜霉菌侵染叶片中的表达模式。序列分析结果表明,Bo CCCH1的基因组全长为1 568 bp,包含1个长度为599 bp的内含子,2个长度分别为627 bp、342 bp的外显子,编码322个氨基酸,蛋白质的分子量与等电点分别为35 296.42 Da和8.47;Bo CCCH1有2个锌指结构,类型均为C-X8-C-X5-C-X3-H。系统发育分析结果表明,Bo CCCH1与其它植物的21条同源序列在进化树上分为6组,来自十字花科的CCCH与Bo CCCH1处于同一分支。基因表达结果表明,Bo CCCH1在叶、花茎、嫩角果、花蕾和花中表达,其中花茎和嫩角果的表达量最高,但在根中未检测到表达;在霜霉菌侵染下,叶片Bo CCCH1的表达量升高,到24 h和36 h时达到最大,推测Bo CCCH1与霜霉病抗性相关。本研究为进一步探明Bo CCCH1在霜霉病抗性反应中的功能奠定了基础。     

NaCl胁迫对白蜡种子生理特性的影响

为了给盐碱地造林树种选择提供依据,以采自秦皇岛和保定的白蜡种子为试材,采用不同浓度的NaCl胁迫处理,研究了其萌发过程中发芽率、细胞膜透性、保护酶活性、丙二醛MDA的变化,结果表明:(1)随着NaCl胁迫浓度的增加,两地白蜡种子发芽率均呈现逐渐降低的变化趋势,秦皇岛各处理的白蜡种子发芽率分别比对照降低了8.9%、52.6%、56.3%、61.6%、87.9%、91.1%,而保定的白蜡种子分别比对照降低20.2%、56.2%、68.7%、86.5%、89.9%、100.0%;(2)随着浓度的升高,种子的相对电导值、K+、Na+渗透率升高,种子的SOD酶活性降低,MDA含量增加,NaCl胁迫浓度越高,种子所受到的伤害越深;(3)0.20%浓度的NaCl胁迫对白蜡种子萌发影响不大。对于不同种源的白蜡种子,秦皇岛的白蜡种子抗盐性要优于保定的白蜡种子。     

Contribution of Different Organs to Grain Filling in Durum Wheat under Mediterranean Conditions I. Contribution of Post‐Anthesis Photosynthesis and Remobilization

Under Mediterranean conditions, drought affects cereals production principally through a limitation of grain filling. In this study, the respective role of post‐anthesis photosynthesis and carbon remobilization and the contribution of flag leaf, stem, chaff and awns to grain filling were evaluated under Mediterranean conditions in durum wheat (Triticum turgidum var. durum) cultivars. For the purpose, we examined the effects of shading and excision of different parts of the plant and compared carbon isotope discrimination (Δ) in dry matter of flag leaf, stem, chaff, awns and grain at maturity and in sap of stem, flag leaf, chaff and awns, this last measurement providing information on photosynthesis during a short period preceding sampling. Source–sink manipulations and isotopic imprints of different organs on final isotope composition of the grain confirmed the high contribution of both carbons assimilated by ears and remobilized from stems to grain filling, and the relatively low contribution of leaves to grain filling. Grain Δ was highly and significantly associated with grain yield across treatments, suggesting the utilization of this trait as an indicator of source–sink manipulations effects on grain yield. Chaff and awns Δ were better correlated with grain Δ than stem and leaf Δ, indicating that chaff were more involved in grain filling than other organs. Moreover, in chaff, sap Δ was highly significantly correlated with dry matter Δ. These results suggest the use of Δ for a rapid and non‐destructive estimation of the variation in the contribution of different organs to grain filling.     

圆柏对几种农田杂草及作物的化感潜力分析

为探究圆柏（Sabina chinensis）的化感作用，本研究采用培养皿滤纸法研究圆柏叶片浸提液对地肤（Kochia scoparia）、稗草（Echinochloa crusgalli）、藜（Chenopodium album）、反枝苋（Amaranthus retroflexus）、马齿苋（Portulaca oleracea）5种杂草种子和向日葵（Helianthus annuus）、燕麦（Avena sativa）、油菜（Brassica campestris）、荞麦（Fagopyrum esculentum）4种作物萌发和生长的影响，并用圆柏叶片、树枝、树皮粉末和浸提液进行盆栽抑草效果检测。结果表明：圆柏叶片浸提液对5种杂草种子萌发和幼苗生长具有显著的抑制作用，对地肤、藜、反枝苋萌发及生长的化感效应指数均在-0.98以上；盆栽抑草结果表明，圆柏不同部位对5种杂草种子萌发和稗草及反枝苋株高生长均有较好的抑制作用，叶片化感强度大于树皮和树枝，叶片粉末拌土处理抑草效果好于浸提液叶面喷施；对作物的化感作用结果表明，圆柏叶片浸提液稀释1.5倍时，抑制荞麦种子萌发但不影响其株高及根生长，不影响燕麦种子萌发但降低其株高和根长生长，抑制油菜的发芽势和发芽指数但不影响其种子萌发率，促进油菜株高及根长生长，不影响向日葵种子萌发和生长。综上所述，圆柏具有良好的抑草活性，对作物影响较小。     

不同温度下TMV侵染枯斑三生烟的LncRNA差异表达

【目的】 筛选不同温度下烟草花叶病毒（Tobacco mosaic virus,TMV）侵染后枯斑三生烟（Nicotiana tabacum var. Samsun NN）差异表达的长链非编码RNA（long non-coding RNA,lncRNA）,研究lncRNA在枯斑三生烟抗性反应中的作用。【方法】 N基因的温度敏感性使枯斑三生烟在25℃时具备对TMV的抗性、在31℃抗性丧失,在这两个温度条件下对枯斑三生烟接种TMV和磷酸盐缓冲盐水（phosphate buffered saline,PBS）,48 h后提取系统叶总RNA,构建链特异性文库后进行深度测序。对测序结果进行过滤后利用HTSeq将有效数据与近缘品种TN90（N. tabacum var. TN90）基因组比对,筛选得到lncRNA后利用FPKM法估计lncRNA的表达水平。通过edgeR筛选差异表达lncRNA（differentially expressed lncRNA,DElncRNA）,并利用qRT-PCR技术对这一结果进行验证。通过共定位及共表达分析预测DElncRNA的靶基因,通过参考基因组注释、GO和KEGG富集分析研究靶基因的功能。【结果】 4个处理共12个样本经lncRNA-seq各测得约8 000万条clean reads,共获得4 737条已知lncRNA、40 169条新lncRNA。其中64个lncRNA在不同温度条件下TMV侵染后存在差异表达,qRT-PCR测定结果显示这些lncRNA的测序正确率在80%左右,表明本研究所得测序数据具备较高的可信度。对DElncRNA进行靶基因预测,发现一些基因同时被25℃下调和31℃上调的DElncRNA靶向。靶基因注释功能丰富,主要参与植物抗病、激素和代谢等生理过程。部分可能与激素通路相关的lncRNA,在25℃下TMV侵染时呈现下调趋势,而在31℃下TMV侵染则呈现上调趋势。GO富集分析显示靶基因主要参与构成膜、囊泡等组分,具备钙、钾离子通道抑制剂活性等分子功能,使相应离子得以转运引发随后的反应,同时也参与发病、抗原加工和呈现、细胞分裂素代谢等生理过程。KEGG分析发现靶基因显著富集在植物激素信号转导通路,25℃下调和31℃上调的DElncRNA靶基因同时富集在激素信号传导、ABC运输蛋白、苯丙烷类生物合成等通路。【结论】 不同温度（25℃和31℃）条件下TMV侵染枯斑三生烟后,长链非编码RNA差异表达,DElncRNA通过作用于激素信号传导、物质转运等过程参与寄主系统获得性抗性反应。研究结果可为揭示植物系统获得性抗性中lncRNA的调控功能以及新型抗病毒技术开发提供依据。     

Effects of cerium on the antioxidant defence system in the petals and the contents of pigments in the calyces of Rosa chinensis Jacq. cut flower

To study the role of cerium (Ce) in regulating the vase life of Rosa chinensis Jacq. cut flowers, the effects of cerium nitrate (Ce(NO₃)₃) on the activities of antioxidant enzymes, the contents of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) in the petals, and the contents of chlorophyll a, chlorophyll b and carotenoids in the calyces were investigated. The results showed that Ce(NO₃)₃ evidently enhanced the activities of peroxidase (POD), catalase (CAT), superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR), glutathione S-transferase (GST) and glutathione peroxidase (GPX), and evidently decreased the contents of MDA and H₂O₂ in the petals, compared with the control. Moreover, Ce(NO₃)₃ evidently increased the contents of chlorophyll a, chlorophyll b and carotenoids in the calyces. The results also showed that Ce(NO₃)₃ evidently increased the numbers of open flowers and decreased the numbers of wilted flowers. Our results suggest that Ce(NO₃)₃ extended the vase life of R. chinensis Jacq. cut flower by improving the antioxidant defence system in the petals and the contents of pigments in the calyces.